Proteomic analyisis of cultured human podocytes stimulated with TNFa - cell pellet
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ABSTRACT: TNFa stimulated human cultured podocytes compared with unstimulated (vehicle control (VC)) podocytes. Incubation of podocytes with high concentrations of TNF alpha led to an overexpression of ICAM1 in particular. We studied this system as a comparator for organoids also stimulated with the same concentration of TNFa. Four conditions (6 replicates each) 1) treatment (5 ng/mL TNFa) for 24 hours 2) control (PBS) for 24h 3) treatment (5 ng/mL TNFa) for 48 hours 4) control (PBS) for 48h
Project description:TNFa stimulated human cultured podocytes compared with unstimulated (vehicle control (VC)) podocytes. Incubation of podocytes with high concentrations of TNF alpha led to an overexpression of ICAM1 in particular. We studied this system as a comparator for organoids also stimulated with the same concentration of TNFa. Four conditions (6 replicates each) 1) treatment (5 ng/mL TNFa) for 24 hours 2) control (PBS) for 24h 3) treatment (5 ng/mL TNFa) for 48 hours 4) control (PBS) for 48h
Project description:We report the use of bulk RNAseq of AML and endothelial cell lines in four conditions (1. Untreated; 2. IFNg - 10 ng/ml; 3. TNFa - 10 ng/ml; 4. IFNg and TNFa - 10 ng/ml) in order to identify genes that are differentially expressed between the two cell types, both at baseline and in an inflammatory microenvironment.
Project description:Calu-3 2B-4 cells were stimulated with interleukin-1a or TNFa. Transcriptional analysis of the extracted RNA was done by microarray. Calu-3 2B-4 cells were washed with phosphate-buffered saline (PBS) and treated with either IL-1a (0.001ng/ml) or TNFa (0.05ng/ml) or mock diluted in PBS for 40 min at 37C. Following treatment, cells were washed 3 times, and fresh medium was added. Triplicate Calu-3 2B4 cultures and triplicate time-matched mock-infected controls were harvested at 3, 6 and 24h for IL-1a and 6 and 24h for TNF post-exposure for transcriptional analysis.
Project description:Wild-type bone marrow-derived macrophages (BMDMs) were treated with vehicle (0.1% EtOH/D-PBS), 6h 100 ng/ml lipopolysaccharide (LPS) or 16h 1uM dexamethasone (Dex) and 6h 100 ng/ml LPS (Dex+LPS) and mRNA expression analysed by RNA-Seq.
Project description:The wild type (WT) and TLR4 -/- pulmonary fibroblasts were treated with phosphate-buffered saline (PBS), 1 µg/mL recombinant mouse (rm) CIRP, 2 ng/mL rmTGF-ß1 (R&D Systems), or rmCIRP plus rmTGF-ß1. The cell lysates collected at 24 hours were used for high throughput mRNA sequencing.
Project description:Balbc J mouse received vehicle (solvent) or OGG1 inhibitor 1 h prior to treatment via intraperitoneal route. Tumor necrosis factor alpha (TNFa, 20 ng per ml) was installed into lungs via intranasal route in 60 microliters of solvent). One hour after TNFa challenge mice were sacrificed, lungs were removed and RNAs were isolated from individual lungs (n=6) After reverse transcription cDNAs were pooled and changes in gene expression was determined by using SABiosciences Mouse Inflammatory Cytokines & Receptors PCR Array (PAMM-011Z)
Project description:Study was carried out to examine how E2 and TNFa together influence gene expression in breast cancer cells compared to either factor alone. Experiment Overall Design: MCF-7 cells were treated with 10 nM E2, 10 ng/ml TNFa, or both for 2h. Experiment was carried out in triplicate.
Project description:Clozapine is an atypical antipsychotic drug for treatment-resistant schizophrenia. Its side effects, including liver enzyme abnormalities, experienced by many patients preclude a more common use as a first-line therapy of schizophrenia. Toxicoproteomics approaches have been demonstrated to effectively guide the identification of toxicological mechanisms. Here, to further our understanding of Clozapine's molecular effects we performed a Data-independent Acquisition (DIA)-based quantitative proteomics investigation of Clozapine-treated human liver spheroid cultures. In total, we quantified 4,479 proteins across five treatment groups (vehicle, 15 µM, 30 µM, 60 µM Clozapine, and 10 ng/mL TNFa + IL-1a). 60 µM Clozapine treatment yielded 36 differentially expressed proteins (fdr < 0.05). Gene-set enrichment analysis indicated perturbation of several gene-sets, which included interferon gamma signaling (e.g., IFNGR1) and prominently autophagy related processes (e.g., upregulation of SQSTM1, MAP1LC3B/LC3B2, GABARAPL2, and NCOA4). Clozapine's effects on autophagy were confirmed using conventional SQSTM1 and LC3B markers by targeted mass-spectrometry and Western Blot.